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ApplicationTimelineStart

SeedlingsTissueHarvest

DNASequencing

QualityControl

GBSPipelineandGSModels

MakeSelection Plant

GenomicSelectiontoIncreasetheRateofGeneticGainofIntermediateWheatgrass(Thinopyrum intermedium)

JaredCrain1,KevinDorn1,TraciKantarski2,JaneGrimwood3,JeremySchmutz3,LeeDeHaan4,JessePoland11KansasStateUniversity,Manhattan,KS66506.2ColumbiaUniversity,NewYorkCity,NY10027.3HudsonAlphaInstituteforBiotechnology,Huntsville,AL35801.4TheLandInstitute,Salina,KS67401.

Malone Family LandPreservation Foundation

Introduction

MaterialsandMethodsCycle6ofTheLandInstituteIWGbreedingprogramformedthe

trainingpopulation,genotyping-by-sequencing(GBS)wasused

formarkerdiscoveryandgenotyping.Thepopulationincluded:

• 3,658genetsinasinglereplicaterow-columndesign.

• Genetswerephenotyped for46traitsovertwoyears.

Seedyield,spikefertility,seedmass,…,stemstrength.

Foreachtrait,1,880-2,400observationswererecorded.

• 2,974plantsgenotypedusingGBS.

Usingtheavailabledata,bestlinearunbiasedpredictors(BLUPs)

werecalculatedforeachtrait.Heritability(h2)wascalculated

usingthegenomicmarkermatrix.Cross-validationproceeded

byleavingentireparentsoutofthetrainingset,topreventbias

ofcloserelativespredictingperformance,andseveralGS

modelsweretestedtoidentifythebestperformingmodels.

ResultsandDiscussion

Refereneces1. DeHaan,L.R.,S.Wang,S.Larson,T.Kantarski,X.Zhang,andD.

Cattani.2014.CurrenteffortstodevelopperennialwheatanddomesticateThinopyrum intermediumasaperennialgrain.In:C.Batello etal.,editors,Perennialcropsforfoodsecurity:Proc.oftheFAOexpertworkshop.p.72–89.

2. Zhang,X.,Sallam,A.,Kantarski,T.,DeHaan,L.R.,…Anderson,J.A.(2016).EstablishmentandOptimizationofGenomicSelectiontoAcceleratetheDomesticationandImprovementofIntermediateWheatgrass.ThePlantGenome,9(1),1–18.

IntermediateWheatgrass(IWG)isapromisingperennialspecies

thatisbeingdomesticatedasagrainandbiomasscrop.Initial

breedingworkbeganattheRodaleInstitute,Kutztown,PA,inthe

1980’s.In2002,TheLandInstitute,Salina,KS,beganbreedingIWG

usingselectionsmadebytheRodaleInstitute[1].Earlyand

contemporaryworkhasbeenencouraging,withseedyield

increasing77%overtwocyclesofselection,andseedmass

increasing23%[1].Whilethesegainshavebeenimportant,ifthe

currentrateofgainsholds,itisestimatedthatitwouldtake

between24and110yearsofbreedingtoreachtheyieldandseed

sizeofannualwheat[1].Therearenumerouschallengesto

breedingIWGincluding:

• Largegenomesize12.6Gb• Allohexaploid(2n=6x=42)• Outcrossingandheterozygous

Asaperennial,breedingIWGisatimeintensiveprocess,withatypicalbreedingcyclelengthoftwoyears(Figure1).

Recently,genomicselection(GS)hasbeenproposedasawayto

increasetherateofgeneticgaininIWG.WorkbyZhangetal.[2]

showedhighpredictiveabilityofGSforseveralagronomictraits,

includingseedweightandbiomass.Inadditiontohighpredictive

ability,GScouldbeusedtodrasticallyreducethelengthofthe

breedingcycle,allowingforintermating ofselectedprogenyeach

year(Figure2)andresultinginabreedingcyclethatistwiceasfast.

GiventhepotentialofGStoincreasetherateofgeneticgain,we

examinedhowGScouldbeappliedanddevelopedmethodsto

applyGStoTheLandInstituteIWGbreedingprogram.

X

Spring2017:Severalhundredcrossesaremadebetweenselectedparents.

X X

… …Summer2018:F1progenyareevaluatedforyieldandagronomictraits.

Spring2019:Thebestplantsfromsummer2018arecrossed,startinganewcycle.

Figure1. TwoyearphenotypicbreedingcycleforIntermediateWheatgrass.

GBSandGSFall2017

1,000plantsthatareplantedinthefieldtotrain

theGSmodel.

100plantswithhighestpredictedvalue,plantedinthegreenhouseforintermating.

Summer2017:UsingF1seedfromspring2017,4,000seedlingsarestarted.

TheseedlingsaregenotypedandpredictedusingGS.Thebestseedlingsformthecrossingblockandalargeportionoflinesgotofieldevaluation.

X XSpring2018:Crossingoccursinthegreenhouse,whilefieldevaluationsprovidedatatoimprovetheGSmodel.

F1seedthatstartsthenextcycleofselection.

Phenotypicevaluation,withphenotypicdatausedtotrainGSmodelforfollowingcycle.

Fall2018:Anewcycleofselectionbegins.

Figure2.ApplicationofgenomicselectiontoIntermediateWheatgrassbreeding.Thegenomicselectionmodelpredictsplantperformance,withthebestplantsformingtheparentsforthenextcycle.Bysimultaneouslyevaluatingmaterialinthefieldandcrossingplants,thetimetocompletethecyclecaneffectivelybecutinhalf.

Table1. Heritabilityandcross-validationpredictionaccuraciesforphenotypictraitsfromCycle6trainingpopulation.Cross-validationaccuracyisreportedasthecorrelationcoefficientbetweenthephenotypicbestlinearunbiasedpredictorandthepredictedvalue.

Trait h2 r2

FreeThreshing2016 0.61 0.45†

FreeThreshing 2017 0.50 0.91†

Spike yieldingrams2017 0.36 0.89Milligramsperseed2016 0.58 0.86Plantheight2016 0.63 0.80

Heritabilityacrossalltraitsrangedfrom0.28-0.67,andthecross-

validationpredictionaccuracywashighforalltraitsrangingfrom

0.45-0.93(Table1).AcrossmultipleGSmodels,theprediction

valuesamonggenetswerequitestable,withtheGaussianKernel

andrrBLUP havingthemostvariationinprediction.Evenwiththis

variation,foranyindividualtraittheminimumcorrelationbetween

thesetwomodelswasr2 >0.94(Figure3).

Aug.1Aug. 25– Sept.5

Sept. 6- 25Sept.26

Sept.27– Oct. 5Oct. 6-11

(byOct.25)

90DaysfromPlantingtoSelection!

Figure3.Comparisonofdifferentgenomicselection(GS)modelsforseedweight2016.ThediagonalliststhedifferentGSmodelsincludingridgeregression,LASSO,andGaussiankernelusingtherrBLUP andBGLRpackages. Thelowertriangleshowsthescatterplotresultsbetweenthemodels,andtheuppertriangledisplaysthecorrelationcoefficientbetweenthedifferentmodels.

rrBLUP_RR

5 6 7 8 9

1.00*** 1.00***−2 −1 0 1 2 3

−2−1

01

2

0.96***

56

78

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● BGLR_RR 1.00*** 0.96***

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● rrBLUP_GAUS

†Furtherworkisinvestigatingthediscrepancybetween2016and2017modelpredictions.

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